Document Type

Poster

Date of Original Version

3-27-2026

Abstract

An accelerated transformation pipeline has been developed for functional genomics and trait gene analysis in Sorghum bicolor based on visual selection using the AmCyan reporter. Here, we combine a GRF4–GIF1 morphogenic regulator with a constitutively expressed AmCyan fluorescent reporter to enable early visual identification of transformed tissues from immature embryo (IE) explants and to shorten time‑to‑regeneration. We demonstrate a practical workflow in which AmCyan‑positive embryogenic clusters are visually recognized and excised 24-48 days after infection (DAI), and then rapidly regenerated directly into plants, in the absence of antibiotic/herbicide selection exposure. Further, we implement a 96‑well microplate reader assay that quantifies AmCyan signal from leaf discs of regenerated plants, enabling a straightforward prescreen prior to confirmatory PCR. AmCyan assessments showed high concordance with PCR results for T‑DNA targets and were orthogonally supported by glufosinate leaf‑paint phenotyping in bar positive plants, indicating the reporter system is a robust proxy for transgene presence. Together, GRF/GIF‑mediated regeneration plus AmCyan‑guided selection provide a streamlined Sorghum transformation pipeline that reduces culture time and handling, supports scalability, and integrates cleanly with standard molecular validation.

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